As is known in the art, confocal microscopy is an imaging technique that can optically image biological objects using confocal microscopes. Confocal microscopes vary in configuration and location of components, but in order to simplify the description, confocal microscopes can be assumed to all function in a similar manner. In a typical confocal microscope, light is directed through a set of holes in a disk, e.g., a Nipkow disk. As the disk spins, the holes that are illuminated produce a scanning pattern similar to that produced by the electron gun of a television tube. Optically, the holes act as pinholes and permit only parallel rays of light to pass. The light exiting the pinholes passes through an objective lens and onto the tissue. Light reflected from the tissue passes back through the objective lens and the pinholes and may be collected by a video camera. The image captured by the video camera can be digitized, computer enhanced, and viewed on a video monitor, stored in a digital or analog form, and/or printed on paper.
The depth to which the confocal microscope can optically penetrate to permit in-vivo observation in real-time is limited by the light penetration into the tissue and the reflective properties of the structures being observed. One skilled in confocal microscopy will recognize that the structures should reflect some light to be visible. Only the light reflected from the biologic structures at the selected plane is allowed to pass into the image plane and to contribute to image formation. Because both the light and the microscope objective lens are focused at the same specific focal plane, objects and structures above and below the plane do not interfere with the formed image.
Attempts to enhance the reflective characteristics of skin tumors and/or lesions typically require that the tumors and/or lesions be excised using Mohs micrographic surgery and then treated for increasing the reflective characteristics thereof. More particularly, Mohs micrographic surgery (MMS) is a surgical procedure based on microscopically controlled excision of cutaneous neoplasms. It offers the highest cure rates among other therapeutic modalities while maximally preserving surrounding normal skin. This therapeutic modality is the treatment of choice for neoplasms in high-risk locations where functional and cosmetic reconstruction is limited, and for histologic tumor subtypes with aggressive biologic behavior. MMS involves excision of the clinically apparent tumor and/or lesion, processing and staining of horizontal frozen sections of the tumor and/or lesion using reagents, such as hematoxylin and eosin or toluidine blue, stepwise microscopic analysis, meticulous mapping of tumor and/or lesion extensions, and re-excision of residual portions of the tumor and/or lesion until tumor-free margins are obtained. Preparation of frozen stained sections of the tumor and/or lesion can require approximately 20–60 minutes per stage. Therefore, Mohs surgery can be a tedious, time-consuming and costly surgical procedure requiring correlating the positive histology with actual skin margins. Although MMS is a tissue-sparing surgical technique, it can still result in the sacrifice of some noninvolved or otherwise healthy tissue.
It would, therefore, be desirable to overcome the aforesaid and other disadvantages.